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Mitochondrial DNA

Methods and Protocols

  • Book
  • © 2009

Overview

Editors:
  1. Jeffrey A. Stuart

    1. Department of Biological Sciences, Brock University, Cpy Catharines, Canada

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  • Covers a wide range of methods for investigating how mitochondrial genetics affect cellular physiology and human health
  • Details unique experimental approaches for studying mtDNA damage and mutation
  • An experimental resource for researchers and physicians concerned with mtDNA disorders
  • Provides an in depth description of tools and approaches used by leading researchers
  • A comprehensive compendium of tools and approaches used in the field

Part of the book series: Methods in Molecular Biology (MIMB, volume 554)

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Table of contents (23 protocols)

  1. Front Matter

    Pages i-xi
    Download chapter PDF

  2. Mitochondrial DNA Replication, Transcription, Translation, and Respiratory Complex Assembly

    1. Front Matter

      Pages 1-1
      Download chapter PDF

    2. Biochemical Isolation of mtDNA Nucleoids from Animal Cells

      • Daniel F. Bogenhagen
      Pages 3-14

    3. Analysis of Mitochondrial DNA by Two-Dimensional Agarose Gel Electrophoresis

      • Aurelio Reyes, Takehiro Yasukawa, Tricia J. Cluett, Ian J. Holt
      Pages 15-35

    4. Comparative Purification Strategies for Drosophila and Human Mitochondrial DNA Replication Proteins: DNA Polymerase γ and Mitochondrial Single-Stranded DNA-Binding Protein

      • Marcos T. Oliveira, Laurie S. Kaguni
      Pages 37-58

    5. Functional Analysis of Mutant Mitochondrial DNA Polymerase Proteins Involved in Human Disease

      • Sherine S. L. Chan, William C. Copeland
      Pages 59-72

    6. Preparation of Human Mitochondrial Single-Stranded DNA-Binding Protein

      • Matthew J. Longley, Leslie A. Smith, William C. Copeland
      Pages 73-85

    7. Methods for Assessing Binding of Mitochondrial Transcription Factor A (TFAM) to DNA

      • Atsushi Fukuoh, Dongchon Kang
      Pages 87-101

    8. Inducible Expression in Human Cells, Purification, and In Vitro Assays for the Mitochondrial DNA Helicase Twinkle

      • Steffi Goffart, Hans Spelbrink
      Pages 103-119

    9. Purification Strategy for Recombinant Formsof the Human Mitochondrial DNA Helicase

      • Tawn D. Ziebarth, Laurie S. Kaguni
      Pages 121-126

    10. Methods for Studying Mitochondrial Transcription Termination with Isolated Components

      • Paola Loguercio Polosa, Stefania Deceglie, Marina Roberti, Maria Nicola Gadaleta, Palmiro Cantatore
      Pages 127-141

    11. Oxidative Phosphorylation: Synthesis of Mitochondrially Encoded Proteins and Assembly of Individual Structural Subunits into Functional Holoenzyme Complexes

      • Scot C. Leary, Florin Sasarman
      Pages 143-162

  3. Mitochondrial DNA Damage and Repair

    1. Front Matter

      Pages 163-163
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    2. Reactive Oxygen Species Production by Mitochondria

      • Adrian J. Lambert, Martin D. Brand
      Pages 165-181

    3. Measuring mtDNA Damage Using a Supercoiling-Sensitive qPCR Approach

      • Sam W. Chan, Junjian Z. Chen
      Pages 183-197

    4. Quantitative Analysis of Oxidized Guanine, 8-Oxoguanine, in Mitochondrial DNA by Immunofluorescence Method

      • Mizuki Ohno, Sugako Oka, Yusaku Nakabeppu
      Pages 199-212

    5. In Vitro Measurement of DNA Base Excision Repair in Isolated Mitochondria

      • Melissa M. Page, Jeffrey A. Stuart
      Pages 213-231

    6. Targeting Repair Proteins to the Mitochondria of Mammalian Cells Through Stable Transfection, Transient Transfection, Viral Transduction, and TAT-Mediated Protein Transduction

      • Christopher A. Koczor, Janet W. Snyder, Inna N. Shokolenko, Allison W. Dobson, Glenn L. Wilson, Susan P. LeDoux
      Pages 233-249

    7. Construction and Characterization of a Cell Line Deficient in Repair of Mitochondrial, but Not Nuclear, Oxidative DNA Damage

      • Sugako Oka, Mizuki Ohno, Yusaku Nakabeppu
      Pages 251-264

  4. Mitochondrial DNA Mutations

    1. Front Matter

      Pages 265-265
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Keywords

About this book

Since the publication of the first edition, the number of unique heritable mtDNA mutations recognized as being associated with bioenergetic dysfunction, cell death and disease has grown. Likewise, our understanding of the basic biology of somatic mtDNA mutations continues to improve. In Mitochondrial DNA: Methods and Protocols, Second Edition, specialists from eight countries share their expertise, providing detailed protocols for studying many aspects of mtDNA. The volume is divided into three sections, which cover the transduction of information from mtDNA to functionally active respiratory complexes, mitochondrial reactive oxygen species (ROS) production as well as mtDNA damage and its repair, and the identification and quantification of heteroplasmic mtDNA mutations. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes sections, highlighting tips on troubleshooting and avoiding known pitfalls.

Comprehensive and authoritative, Mitrochondrial DNA: Methods and Protocols, Second Edition promises to aid researchers in further expanding our knowledge of this vital area of cell biology.

Reviews

From the reviews of the second edition: “The book contains detailed descriptions both of established techniques that continue to be usefully applied and recent information that improves understanding of mtDNA biology. It is intended for graduate students, postdoctoral fellows, and established investigators. I think even teachers in this field and early career investigators will find this book quite useful.” (Omer Iqbal, Doody’s Review Service, January, 2010)

Editors and Affiliations

  • Department of Biological Sciences, Brock University, Cpy Catharines, Canada

    Jeffrey A. Stuart

Bibliographic Information

  • Book Title: Mitochondrial DNA

  • Book Subtitle: Methods and Protocols

  • Editors: Jeffrey A. Stuart

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-59745-521-3

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Humana Press 2009

  • Hardcover ISBN: 978-1-934115-60-2Published: 21 July 2009

  • Softcover ISBN: 978-1-61779-495-7Published: 06 December 2011

  • eBook ISBN: 978-1-59745-521-3Published: 10 June 2009

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 2

  • Number of Pages: XII, 404

  • Number of Illustrations: 73 b/w illustrations, 1 illustrations in colour

  • Topics: Cell Biology, Human Genetics

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